Int. J. Morphol., 28(4):1315-1322, 2010.

Effect of Prolonged Oral Administration of Aqueous Ficus sycomorus Stem-bark Extract on Testicular Size of Growing Albino Rat

 

Efecto de la Administración Oral Prolongada de Extracto Acuoso de la Corteza de Ficus sycomorus en el Tamaño Testicular de Rata Albina en Crecimiento

 

*N. A. Igbokwe; **I. O. Igbokwe & *U. K. Sandabe

* Department of Veterinary Physiology, Pharmacology and Biochemistry, Faculty of Veterinary Medicine, University of Maiduguri, Nigeria.

** Department of Veterinary Pathology, Faculty of Veterinary Medicine, University of Maiduguri, Nigeria.

Correspondence to:

---

SUMMARY: Decoction of Ficus sycomorus stem bark is used to treat infertility involving low sperm counts, but the reproductive risk associated with its use is uncertain. The aqueous stem-bark extract was orally administered at 200-600 mg/kg for 30 days to post-pubertal growing albino rats. Within the period, the live body weights, scrotal diameters and testicular size parameters were estimated. The mean live body weight, scrotal diameter of the control and treated rats significantly (p<0.05) increased as the animals grew, but the treatment did not significantly (p>0.05) affect the parameters. There were no significant (p>0.05) variations in mean testicular size parameters in treated and control rats, and no dose-dependent nor time-dependent effects occurred. Sections of the testes of the treated animals showed active spermatogenesis in the seminiferous tubules with no apparent histological lesions. It is concluded that the use of the crude extract of the Ficus sycomorus within our dose range portends minimal male reproductive risk and underscores the need to study its pro-spermatic effect.

Key words: Ficus sycomorus extract; Testicular size; Testicular histopathology; Spermatogenesis; Male reproductive risk.

---

RESUMEN: La decocción de la corteza del tallo de Ficus sycomorus se usa para tratar la infertilidad relacionada al bajo conteo de espermatozoides, pero es incierto el riesgo reproductivo asociado con su uso. El extracto acuoso de la corteza se administró, en ratas albinas, por vía oral, en una dosis de 200-600 mg/kg, durante 30 días post-crecimiento puberal. Durante el período, fueron controlados los siguientes parámetros: peso corporal in vivo, diámetro escrotal y tamaño testicular. El promedio de peso corporal in vivo y diámetro escrotal de las ratas control y tratadas, tuvieron un incremento significativo (p<0,05) en los animales, pero el tratamiento no afectó significativamente (p> 0,05) esos parámetros. No hubo variaciones significativas (p> 0,05) en la media de los parámetros tamaño testicular en las ratas tratadas y control, ni tampoco se produjeron efectos dependientes de la dosis, ni en función del tiempo. Las secciones de los testículos de los animales tratados mostraron una espermatogénesis activa a nivel de los túbulos seminíferos, sin lesiones histológicas aparentes. Se concluye, que el uso del extracto crudo de Ficus sycomorus, dentro de una dosis de 200-600 mg/kg, presenta mínimo riesgo en la reproducción masculina, sin embargo, se hace necsario estudiar su efecto pro-espermático.

PALABRAS CLAVE: Extracto de Ficus sycomorus; Tamaño testicular; Histopatología testicular; Espermatogénesis; Riesgo reproductivo macho.

---

INTRODUCTION

Ficus sycomorus, a medicinal plant belonging to the class Moracea, is used in Nigeria, Niger, Mali, South Africa, Guinea, Kenya, Tanzania, Somalia, Ethiopia and Ivory Coast as extracts of fruits, leaves, root and stem barks to treat various ailments such as cough, diarrhea, skin infections, stomach disorders, liver disease, epilepsy, tuberculosis, lactation disorders, helminthiasis, infertility and sterility (Palayer, 1977; Heine & Brenzinger, 1988; Gachathi, 1989; Hedberg & Staugard, 1989; Heine & Heine, 1989; Abdurrahman, 1992; Samuelsson et al., 1992; Bekalo et al., 1996; Arnold & Galamain, 2002; Sandabe, 2002; Pakia & Cooke, 2003; Kone, 2005; Moshi-Mainen et al., 2005; Maregesi et al., 2007; AFLORA, 2008; Kone & Atindehou, 2008; Mahenge, 2008; Malgras, 2008). The aqueous extract of the plant had relative safety with LD50 of 720mg/kg, contained pharmacologically active substances such as gallic tannis, saponins, reducing sugars, alkaloids and flavone aglycones, and caused no haematological, hepatic and renal toxicities (Sandabe). The aqueous extract had some anticonvulsive effect as well as inhibitory effect on muscle contractions (Sandabe et al., 2006). Similarly, other extracts of a related plant species, Ficus plantyphylla had sedative, anticonvulsant, muscle relaxant and anesthetic properties (Bokko, 1998; Onyeyili et al., 1998). Because of these nervous system effects, the extracts were used in the management of psychiatric patients (Abdurrahman).

Medicinal plant extract from Carica papaya (Lohiya et al., 1994), Quassia amara (Raji & Bolarinwa, 1997), Terminalia catappa (Ratnasooriya & Dharmasiri, 2000), Ricinus communis (Raji et al., 2006), Garcinia kola (Akpantah et al., 2003), Ruta graveolens (Khouri & El-Akawi, 2005), Alstonia beonei (Raji et al., 2005), Vernonia amygdalina (Oyeyimi et al., 2008) were reported to have adverse effects on male reproduction by reducing the weight of testes. Since Ficus sycomorus had been associated with treatment of fertility and sterility in humans (Pakia & Cooke, 2003; Malgras, 2008) and the effects of its extracts on male reproductive organs had not been studied, the present research was designed to assess the effects of the aqueous extract of the stem bark on the testicular size of growing albino rat.

Material and Method

Animals. Sixty post-pubertal male albino rats, above 50 days of age and weighing 98.0±22.9g, were sourced from Ayuba Animal Farm, Maiduguri 14 days earlier than the time of commencement of the experiment. They were housed in stan dard plastic cages containing 5 rats each with comparable mean live body weights and kept in a room with ambient temperature not exceeding 30C. They were fed ad libitum with commercial pelletized feed (Vital Feed, Jos). Clean drinking water was provided freely in nipple drinkers.

Plant identification and extraction. The tree of Ficus sycomorus was identified by a taxonomist in the Department of Biological Sciences, Faculty of Science University of Maiduguri. The stem bark was chopped off and air dried away from direct sunlight for eight days. The dried stem bark was pounded into powdered form using pestle and mortar. The powdered extract (550g) was dissolved in distilled water (3000ml) in a round bottom flask. The mixture was refluxed for 2 hours and filtered while hot with No.1Watman filter paper. The refluxing was done three times using fresh distilled water each time. After filtration, the filtrate was poured on a tray and placed in a hot air oven at 40-50C for 2 hours for the water to evaporate. The dried sample left on the tray was weighed and used for the experiment.

Experimental design. The rats were divided into four groups of 15 rats each. The concentration of the extract used was 200 mg/ml (Sandabe). Rats in groups A, B and C were administered orally through an esophago-gastric tube (Onu et al., 2008) with 200 mg/kg, 400 mg/kg and 600 mg/kg, respectively, of the extract for 30 days. Rats in group D served as control and received distilled water only during the period. Five rats from each group were humanely sacrificed on days 10, 20 and 30 of treatment to determine the dose-dependent and time-dependent effects of the treatment on body weight gain, scrotal diameter and testicular size. The live body weights and scrotal diameters of the rats were measured. The testes were dissected out and separated from the spermatic cord by cutting the vascular cone. The testes were weighed without the epididymides and the longitudinal length and mid-circumference of the testes were estimated.

Histopathology. One of the testes of each rat was fixed in Bouin's fluid, processed, cut and stained with haematoxylin and eosin (Akpantah et al.).

Statistical analysis. The data obtained is presented as means ± standard deviations and variations in means were assessed by one-way analysis of variance (ANOVA); coefficient of determination, r2, for linear trends in relation to time and dose were determined using computer software (GraphPad Instat, 1993 version, http:// www.graphpadinstat.com ).

Results

Body weight and scrotal diameter.The mean live body weights, weight gain, scrotal diameter and increase in scrotal diameter of the control and treated rats significantly (P<0.05) increased during the time of treatment, but the dose of administration of the extract did not significantly (P>0.05) affect the parameters (Figs. 1 and 2). The coefficient of determination, r2, assessing time-dependent linear trend of the parameters (Table I) were significant (P<0.01) and comparable at various doses, but no significant (P>0.05) dose-dependent linear trend were observed with all the parameters. The time-dependent linear trend was stronger for the percentage increase in scrotal diameter over time (r2=0.36-0.56) than the scrotal diameter per se (r2=0.09-0.17).

Fig. 1. Live body weights and weight gain of albino
rats orally administered with aqueous extract of Ficus
sycomorus stem bark for 30 days.

Fig. 2. Scrotal diameter and percentage increase in the
diameter from day 0 of treatment in albino rats orally administered
with aqueous extract of Ficus sycomorus
stem bark for 30 days.

Testicular size. The effect of oral administration of aqueous Ficus sycomorus stem-bark extract for 30 days on the testicular weight, length, mid-circumference and size index of the rats are summarized in Tables II and III. The extract at various doses did not cause any significant (P>0.05) variations in testicular size parameters during the period of treatment when compared with the untreated control rats, and no dose-dependent nor time-dependent effects occurred with these parameters.

 

 

Testicular morphology. The testes of the treated rats had similar histological appearance as the control rat (Figs. 3 and 4), and showed active spermatogenesis.

Fig. 3. Photomicrograph of the
testis of a control rat (H&E, x 400)

Fig. 4. Photomicrograph of the testis of a rat treated
with Ficus sycomorus extract
at 600 mg/kg for 30 days.

 

Discussion

Aqueous extracts of Ficus sycomorus stem-bark, roots, leaves and fruits have been reportedly used to treat infertility and sterility in humans and animals in parts of Africa (Pakia & Cooke; Kone & Atindehou). The oral administration of aqueous extract of Ficus sycomorus stem-bark at 200-600mg/kg to male albino rats for 30 days did not affect body weights, scrotal diameter and testicular size parameters. The increasing body weights of the growing rats correlated with their testicular size parameters and scrotal diameters (Igbokwe, 2010), but the means of the testicular size parameters marginally rose over the period, with neither significant changes in their means nor linear trend. Mean scrotal diameter varied significantly without the testicular size parameters in the growing rats, indicating that the scrotum housing the testes was affected more by growth changes than testicular size parameters. It was expected that any effect of the extract administration would influence the scrotal diameter more than the testicular size parameters, having shown that the scrotal diameters directly correlated with testicular size parameters (Igbokwe). Testicular weights are relatively stable and vary minimally within a given species and change in testicular size parameter is an outcome of a significant alteration in structure of the testis (Collins et al., 2006).

The morphology of the testes of the rats was not adversely affected by the treatment, an observation that corroborated the lack of decrease in the testicular size parameters. This indicated that the extract had little or no toxicity for the testes. Several plant extracts that caused histological damage of the seminiferous tubules invariably elicited decline in testicular size parameters and were accompanied by reduced function of the testes (Lohiya et al.; Raji & Bolarinwa; Ratnasooriya & Dharmasiri; Akpatah et al.; Khouri & El-Akawi, 2005; Raji et al., 2005; Raji et al., 2006; Oyeyimi et al.). On the other hand, extract of Lepidium meyenii (Maca) increased sperm production without change in testicular size (Chung et al., 2005). At the present dose of administration of Ficus sycomorus extract, the seminiferous tubular epithelium of the testis was not depleted and there was evidence of active spermatogenesis. The extract increased testicular and epididymal sperm cell counts on day 30 of the treatment (Igbokwe), suggesting a pro-spermatic effect on the spermatogenic stages of the germinal epithelium. Thus, those who use decoctions of Ficus sycomorus stem bark at 200-600 mg/kg for treatment of low sperm count may be exposed to minimal reproductive risk.

In conclusion, the oral administration of aqueous extract of Ficus sycomorus stem bark did not affect testicular size parameters and the related scrotal diameters of rats, because it caused no obvious histopathological change in the epithelium of the seminiferous tubules.

Acknowledgements

Our appreciations go to Mr. Fine Akawo for the extraction of the stem bark in the Department of Chemistry, University of Maiduguri, Maiduguri. The project was privately funded by one of the authors under the Strategic Animal Research Initiative, Nigeria.

 

References

Abdurraham, F. Studies on natural products chemistry of the Moraceae in African traditional medicine and management of psychiatry in Borno state. MSc dissertation, University of Maiduguri, Maiduguri, 1992.

AFLORA. The data base of traditional plant utilization in Africa. Center for African Area studies, Kyoto University. Accessed 28th October 2008. Available in: http//www.130.54.103.36/aflora.nsf.

Akpantah, A. O.; Oremosu, A. A.; Ajala, M.; Noronha, C. C. & Okanlawon, C. C. The effect of crude extract of garcina kola seed on the histology and hormonal milieu of male Sprague Dawley rats' reproductive organs. Nig. J. Health Biomed. Sciences, 2:40-6, 2003.

Arnold, H. H. & Gulamain, M. Pharmacopia of traditional medicine in Venda. J. Ethnopharmacol., 12:35-74, 2002.

Bekalo, I.; Keengwe, M.; Mathins, E. & Mundy, P. ITDG and IIRR. Ethnoveterinary medicine in Kenya. A field Manual of Traditional Animal Care Practice. Intermediate Technology Development Group and International Institute of Rural Construction, Nairobi Kenya, 1996. p.266

Bokko, P. M. Effects of Ficus platyphylla stem bark extract on ketamine induced anesthesia in rabbits. MSc dissertation, University of Maiduguri, Maiduguri, 1998.

Collins, T. F. X.; Sprando, R. L.; Shackleford, M. T.; Gruber, M. F. & Morse, D. E. Principles of risk assessment- FDA perspective. In: Hood, R. D. (Ed.). Developmental and reproductive toxicology-a practical approach. 2 ed. Boca Raton, Florida, CRC Press, 2006. pp.877-910.

Chung, F.; Rubio, J.; Gonzales, C.; Gasco, M. F. & Gonzales, G. F. Dose-response effects of Lepidium meyenii (Maca) aqueous extract on testicular function and weight of different organs in adult rats. J. Ethnopharmacol., 98:143-7, 2005.

Gachathi, F. N. Kikuyu botanical dictionary of plant names and uses. Nairobi, AMREF, 1989.

Hedberg, I. & Staugard, F. Traditional medicine in Botswana, traditional medicinal plants. The Nordic School of Public Health. Stockholm, Ipelegeng Publishers, 1989. pp.324.

Heine, B. & Brenzinger, J. Plant concepts and plant use. An ethnobotanical survey of the semi-arid and arid lands of East Africa. Part 4: Plant of Boran (Ethiopia and Kenya). B and G. Saarbrucken, Verlag Breintenbach Publishers, 1988. pp.26.

Heine, B. & Heine, I. Plant concepts and plant use. An ethnobotanical survey of the semi-arid and arid lands of East Africa. Part 1: Plants of the Chamus (Kenya), B and G. Saarbrucken, Verlag Breintenbach Publishers, 1989. pp.104.

Igbokwe, N. A. Effect of aqueous extract of Ficus sycomorus stem bark on sperm cell production in male albino rats. MSc Dissertation, University of Maiduguri, Maiduguri, 2010.

Khouri, N. A. & El-Akawi, Z. Antiandrogenic activity of Ruta graveolens L in male Albino rats with emphasis on sexual and aggressive behavior. Neuro. Endocrinol. Lett., 26:823-9, 2005.

Kone, W. M. Uses of 25 plant species as ethnoveterinary medicinal plants. Journal of Botany, 2005. Accessed 28th October 2008. Avalaible in: www. Linkinghub.elsevier.com/retrieve/pii/ so254629907003651

Kone, W. M. & Atindehou, K. K. Ethnobotanical inventory of medicine in northern Cote d'Ivore (West Africa). South African Journal of Botany, 74:76-84, 2008.

Lohiya, N. K.; Goyal, P. B.; Jayaprakash, D.; Ansari, A. S. & Sharma, S. Antifertility effects of aqueous extract of Carica papaya seeds in male rats. Planta Med., 60:400-4, 1994.

Mahenge, C. P. I. Ficus sycomorus, 2008. Accessed 28th October 2008. Avalaible in: www.easternarc.or.tz/downloads/mahenge/FT/20%kr%20NTP%20report

Malgras, D. Prelude medicinal plants database, metafro, 2008. Accessed October 28th 2008. Avalaible in: www.metafro.be/prelude/view_symptom

Maregesi, S. M.; Ngassapa, O. D.; Pieters, L. & Vlietinck, A. J. Ethnopharmacological survey of the Bunda district, Tanzania. Plants used to treat infectious diseases. J. Ethnopharmacol., 113:457-70, 2007.

Moshi-Mainen, J.; Godeliver, A.B.; Kagashe, I. & Mbwambo, Z. H. Plants used to treat epilepsy by Tanzanian traditional healers. J. Ethnopharmacol., 97:327-36, 2005.

Onu, J. E.; Oke, B. O.; Ozegbe, P. C. & Oyewale, J. O. Effects of alcohol on testicular weight and testosterone levels in Wistar rats exposed during gestation and lactation. Proceedings of the 44th Annual Congress of the Nigerian Veterinary Medical Association, Delta 2007, pp. 116-121, 2008.

Onyeyili, P. A.; Akiniyi, I. A. & Yakubu, T. U. Sedative, anticonvulsant and muscle relaxant effect of Ficus plantyphilla stem bark extract in rats. West African J. Biol. Sci., 7:161-8, 1998.

Oyeyimi, M. O.; Ajala, O. O.; Leigh, O. O. & Adesiji, T. F. The spermiogram of Wistar rats treated with aqueous leaf extract of Vernonia amygdalina. Proceedings of the 44th annual congress of the Nigerian Veterinary Medical Association, Delta 2007, pp.89-92, 2008.

Pakia, M. & Cooke, J. A. The ethnobotany of Midzichenda tribes of the coastal forest areas in Kenya: 2. Medicinal plant uses. South African Journal of Botany, 69:328-95, 2003.

Palayer, P. Lexique des plantes du payssar plantes spontannes et cultivees. Tome 2. College Charles Lwnngi C.E.L. Sahr-TChad. Publie avec le concours financier de l'UNESCO, 1977. pp.78

Raji, Y. & Bolarinwa, A. F. Antifertility activity of Quassia amara in male rats-in vivo study. Life Sci., 61:1007-14, 1997.

Raji, Y.; Salman, T. M. & Akinsemisoye, O. S. Reproductive functions in male rats treated with methanolic extract of Alstonia beonei stem bark. Afr. J. Biomed. Res., 8:105-11, 2005.

Raji,Y.; Oloyo, A. K. & Morakinyo, A. O. Effect of methanol extract of Ricinus communis seed on reproduction of male rats. Asian J. Androl., 8:105-11, 2006.

Ratnasooriya, W. D. & Dharmasiri, M. G. Effects of Terminalia catappa seeds on sexual behaviour and fertility of male rats. Asian J. Androl., 8:115-21, 2000.

Samuelsson, G. M.; Faraha, H.; Claeson, P.; Hages, M.; Thulin, M.; Hedberg, O.; et al. Inventory of plants used in traditional medicine in Somalia III. Plants of the families Lauracae-papilionaceae. J. Ethnopharmacol., 37:93-112, 1992.

Sandabe, U. K. Phytochemical and toxicological studies of aqueous extract of Ficus sycomorus in laboratory animals. PhD thesis, University of Maiduguri, Maiduguri, 2002.

Sandabe, U. K.; Onyeyili, P. A. & Chibuzo, G. A. Phytochemical screening and effect of aqueous extract of Ficus sycomorus L (moraceae) stem bark on muscular activity in laboratory animals. J. Ethnopharmacol., 103:481-3, 2006.

N. A. Igbokwe
Department of Veterinary Physiology
Pharmacology and Biochemistry,
University of Maiduguri
P. M. B. 1069
Maiduguri, NIGERIA

Tel: + 234 8060175771

Email: nanaigb@yahoo.com

naigbokwe@gmail.com

Received: 20-05-2009
Accepted: 04-11-2009